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Short-term toxicity to fish

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Reference
Endpoint:
short-term toxicity to fish
Type of information:
experimental study
Adequacy of study:
key study
Study period:
13 August 2002 to 7 August 2003
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to
Guideline:
OECD Guideline 203 (Fish, Acute Toxicity Test)
Deviations:
not specified
Qualifier:
according to
Guideline:
EU Method C.1 (Acute Toxicity for Fish)
Deviations:
not specified
Principles of method if other than guideline:
The test was carried out in accordance to the protocol entitled: "Semi-static acute toxicity test with the zebra fish Brachydanio rerio", which was approved by the study director on 13 August 2002.
GLP compliance:
yes
Analytical monitoring:
yes
Details on sampling:
No information provided.
Vehicle:
no
Details on test solutions:
The test solution was dissolved directly into the dilution water; the dilution water was also used as the control.
Test organisms (species):
Danio rerio (previous name: Brachydanio rerio)
Details on test organisms:
The average total length and weight (+/- standard deviation) of 10 fish of the batch used (randomly taken at the start of the test) were 2.8 +/- 0.14 cm and 0.18 +/- 0.02 g, respectively. The fish were cultured under the circumstances of the test.
Test type:
semi-static
Water media type:
freshwater
Limit test:
no
Total exposure duration:
96 h
Post exposure observation period:
Not applicable.
Hardness:
Not stated.
Test temperature:
24 ± 1 °C (variation of temperature measured in one of the control vessels: 24-24.7 ºC)
pH:
6.6-8.2
Dissolved oxygen:
Lowest measured oxygen concentration: 7.8 mg O2/L
Salinity:
Not applicable.
Nominal and measured concentrations:
10, 18, 32, 56, 100 mg/L (nominal)
Details on test conditions:
- The test was performed in 3.2 litre all-glass aquaria, each containing 3.0 litre of exposure medium; the aquaria were covered with a perspex plate. One aquarium containing 10 fish was used for each exposure concentration (one replicate per concentration). The fish were not fed 24h prior to the test and during the test.

- The test vessels were coded with the study number and with the test material concentration. The fish were added randomly, using a random table. The exposure media were slightly aerated from t=3 h and replaced every 48h.

- At the beginning and at the end of the test, as well as at every 24 h the pH and the oxygen concentration of the exposure mediua were measured. At t=48 h both the spent media, just before replacement, and the newly prepared media, just after dosing were measured. At the same times the temperature in the clean and spent control vessel was measured and should be 24 ± 1 °C.

- The dead and living animals were counted and if applicable the dead ones removed after 3, 24, 48, 72, and 96 hours. At the same time the condition (swimming behaviour, colour or any other visually observable morphological or behavioural criterion) of the survisors was compared wih that of the control animals.

- The actual concentrations of the test material in the exposure media were determined by chemical analysis.

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: The dilution water used was DSWL-E prepared from ground water. DSWL-E has proven to be suitable for the culture of the zebra fish Brachydanio rerio.

OTHER TEST CONDITIONS
- Photoperiod: 16h light/8h dark
Reference substance (positive control):
no
Key result
Duration:
3 h
Dose descriptor:
LC50
Effect conc.:
> 100 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Key result
Duration:
24 h
Dose descriptor:
LC50
Effect conc.:
> 100 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Key result
Duration:
48 h
Dose descriptor:
LC50
Effect conc.:
> 100 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Key result
Duration:
72 h
Dose descriptor:
LC50
Effect conc.:
> 100 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Key result
Duration:
96 h
Dose descriptor:
LC50
Effect conc.:
> 100 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Key result
Duration:
96 h
Dose descriptor:
LC100
Effect conc.:
> 100 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Key result
Duration:
96 h
Dose descriptor:
NOEC
Effect conc.:
>= 100 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Key result
Duration:
96 h
Dose descriptor:
NOEC
Effect conc.:
56 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
other: condition
Details on results:
OBSERVATIONS ON THE EXPOSURE MEDIA
The 0, 10 and 18 mg/L solutions were clear and colourless. At the higher concentrations, the media where somewhat turbid. At the end of the test (t = 48 h), flocks were observed on the bottom of the test vessels in the old (spent) solutions of 32, 56 and 100 mg/L. At the end of the test (t = 96h), the solutions of 0 and 18 mg/L were clear, at 32 mg/L a slight turbidity was observed; at 56 and 100 mg/L the solutions were somewhat milky and at 100 mg/L there was some precipitation.

CHEMICAL ANALYSIS
At t = 0 h and t = 48 h, just after dosing, the average measured concentrations were 97.9 and 97.1 % of the nominal concentration respectively. At t = 48h and t = 96 h, after 48h exposure, the measured concentration in the same media was 96.9 and 97.3 % respectively. As this is above 80 % of the nominal concentration, the results of the test are expressed in nominal concentrations.

SURVIVAL AND OTHER OBSERVATIONS ON THE TEST FISH
At the end of the test, all fish were alive in the control and in the exposure media. However, at 100 mg/L, slow swimming fish, fish swimming higher in the test vessel and some fish showing a lighter colour were observed. It is therefore concluded that 100 mg/L is the NOEC for survival and 56 mg/L is the NOEC for the conditional endpoint.
Results with reference substance (positive control):
not applicable.
Reported statistics and error estimates:
LC50, LC100 and NOEC values were determined.
Validity criteria fulfilled:
yes
Conclusions:
Under the circumstances of the test, the 96h LC50 of the test material for the zebra fish Brachydanio rerio is > 100 mg/L and the 96h NOEC for mortality >=100 mg/L.
Executive summary:

The acute toxicity of the test material to the fresh-water fish species Brachydanio rerio was determined in line with OCED Guideline 203, under GLP conditions. The duration of the test was 96 hours.

The test was carried out under semi-static conditions with replacement of the exposure media at t = 48 h and with 10 fish for the control medium and for each test material concentration. The nominal concentrations tested were 10, 18, 32, 56 and 100 mg/L. The test material was dissolved directly in the dilution water. The dilution water was also used as control medium.

During the test some flocks, turbidity and precipitation were observed at the test material concentrations of 32 mg/L and higher. The control and the 10 and 18 mg/L solutions were clear throughout the test. The actual concentration of the test material in the exposure media were determined by chemical analysis. At t = 0 h and t = 48 h, just after dosing, the average measured concentrations were 97.9 and 97.1 % of the nominal concentration respectively. At t = 48h and t = 96h, after 48h exposure, the measured concentration in the same media was 96.4 and 97.3 % respectively. As this is above 80 % of the nominal concentration, the results of the test are expressed as nominal concentrations.

There was no mortality observed throughout the test, however at 100 mg/l, the fish showed no adverse effects with respect to condition.

Under the circumstances of the test, the 96h LC50 of the test material for the zebra fish Brachydanio rerio is > 100 mg/L and the 96h NOEC for mortality >=100 mg/L.

Description of key information

Key Study: Hooftman & de Wolf (2003/2004)

Under the circumstances of the test, the 96h LC50 of the test material for the zebra fish Brachydanio rerio is > 100 mg/L and the 96h NOEC for mortality >=100 mg/L.

Key value for chemical safety assessment

LC50 for freshwater fish:
100 mg/L

Additional information

The key study presented for this endpoint and used for classification and labelling purposes is Hooftman & de Wolf, 2003 as amended by Hooftman & de Wolf, 2004.

The acute toxicity of the test material to the fresh-water fish species Brachydanio rerio was determined in line with OCED Guideline 203, under GLP conditions. The duration of the test was 96 hours.

The test was carried out under semi-static conditions with replacement of the exposure media at t = 48 h and with 10 fish for the control medium and for each test material concentration. The nominal concentrations tested were 10, 18, 32, 56 and 100 mg/L. The test material was dissolved directly in the dilution water. The dilution water was also used as control medium.

During the test some flocks, turbidity and precipitation were observed at the test material concentrations of 32 mg/L and higher. The control and the 10 and 18 mg/L solutions were clear throughout the test. The actual concentration of the test material in the exposure media were determined by chemical analysis. At t = 0 h and t = 48 h, just after dosing, the average measured concentrations were 97.9 and 97.1 % of the nominal concentration respectively. At t = 48h and t = 96h, after 48h exposure, the measured concentration in the same media was 96.4 and 97.3 % respectively. As this is above 80 % of the nominal concentration, the results of the test are expressed as nominal concentrations.

There was no mortality observed throughout the test, however at 100 mg/l, the fish showed no adverse effects with respect to condition.

Under the circumstances of the test, the 96h LC50 of the test material for the zebra fish Brachydanio rerio is > 100 mg/L and the 96h NOEC for mortality >=100 mg/L.